Free-flying honeybees exhibit remarkable cognitive capacities but the neural underpinnings of these capacities cannot be studied in flying insects. Conversely, immobilized bees are accessible to neurobiological investigation but display poor visual learning. To overcome this limitation, we aimed at establishing a controlled visual environment in which tethered bees walking on a spherical treadmill learn to discriminate visual stimuli video projected in front of them. Freely flying bees trained to walk into a miniature Y-maze displaying these stimuli in a dark environment learned the visual discrimination efficiently when one of them (CS+) was paired with sucrose and the other with quinine solution (CS−). Adapting this discrimination to the treadmill paradigm with a tethered, walking bee was successful as bees exhibited robust discrimination and preferred the CS+ to the CS− after training. As learning was better in the maze, movement freedom, active vision and behavioral context might be important for visual learning. The nature of the punishment associated with the CS− also affects learning as quinine and distilled water enhanced the proportion of learners. Thus, visual learning is amenable to a controlled environment in which tethered bees learn visual stimuli, a result that is important for future neurobiological studies in virtual reality.

Central-place foragers exploiting floral resources often use multidestination routes (traplines) to maximise their foraging efficiency. Recent studies on bumblebees have showed how solitary foragers can learn traplines, minimising travel costs between multiple replenishing feeding locations. Here we demonstrate a similar routing strategy in the honeybee (Apis mellifera), a major pollinator known to recruit nestmates to discovered food resources. Individual honeybees trained to collect sucrose solution from four artificial flowers arranged within 10 m of the hive location developed repeatable visitation sequences both in the laboratory and in the field. A 10-fold increase of between-flower distances considerably intensified this routing behaviour, with bees establishing more stable and more efficient routes at larger spatial scales. In these advanced social insects, trapline foraging may complement cooperative foraging for exploiting food resources near the hive (where dance recruitment is not used) or when resources are not large enough to sustain multiple foragers at once.